New Generation Sequencing (NGS) technologies and qRT-PCR associated with specific bioinformatics programs aim to discover and validate new DNA/RNA/genomics/transcriptomics biomarkers. The objective: developing new diagnostics in veterinary applications .
Gene expression profiles (transcriptomes) through sequencing technologies (New Generation Sequencing, NGS) and specific bioinformatics tools aim to the acquisition of new DNA/RNA/genomics/transcriptomics data. This quantitative process helps for identifying all the biomarkers involved in each physiopathological situation in an exhaustive and sensitive manner. However, the genes are either already known or not, the genome of the organism completely sequenced or not. .
But, specific bioinformatics programs and database are fully required for analyzing the results generated by these New Generation Sequencing (NGS) systems, including sequence analysis, gene annotation, computational evolutionary biology, measuring biodiversity, analysis of regulation, analysis of protein expression, analysis of mutation, etc .
After these bioinformatics treatments, all the results can be implemented in an interactive database and can be compared with other gene expression profiles (transcriptomes) established through sequencing. Thus, each user can easily search and compare gene expression profiles (transcriptomes), and can select sets of specific biomarkers dedicated to a tissue or a pathology ...
To validate these biomarkers or in a high throughput routine, quantitative Real-Time PCR can be used for monitoring global patterns of gene expression. This technique provides an absolute quantitative analysis of gene expression and a rapid and precise confirmation of genetic changes.
Quantitative Real-Time PCR:
Is the most sensitive technique for mRNA detection and quantification ,
Can be used to quantify mRNA levels from much smaller samples ,
Gives more accurate measurements and is much better adapted to analyses performed on
large numbers of samples ,
Has been successfully developed and has become a reliable and easy to use standard method
for the detection and/or quantification of nucleic acid sequences ,
Reduces hands-on time and increases reliability ,
Has been adopted for a wide range of new applications .
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